The selectin GMP-140 binds to sialylated, fucosylated lactosaminoglycans on both myeloid and nonmyeloid cells.

Granule membrane protein-140 (GMP-140) is an inducible receptor for myeloid leukocytes on activated platelets and endothelium. Like other selectins, GMP-140 recognizes specific oligosaccharide ligands. However, prior data on the nature of these ligands are contradictory. We investigated the structural features required for ligand interaction with GMP-140 using purified GMP-140, cells naturally expressing specific oligosaccharides, and cells expressing cloned glycosyltransferases. Like the related selectin endothelial leukocyte adhesion molecule-1 (ELAM-1), GMP-140 recognizes alpha(2-3)sialylated, alpha(1-3)fucosylated lactosaminoglycans on both myeloid and nonmyeloid cells, including the sequence Neu5Ac alpha 2-3Gal beta 1-4(Fuc alpha 1-3)GlcNac beta-R (sialyl Lewis x). Recognition requires sialic acid, because cells expressing large amounts of Lewis x, but not sialyl Lewis x, do not interact with GMP-140. Although sialyl Lewis x is expressed by both myeloid HL-60 cells and CHO cells transfected with an alpha 1-3/4 fucosyltransferase, GMP-140 binds with significantly higher affinity to HL-60 cells. Thus, the sialyl Lewis x tetrasaccharide may require additional structural modifications or specific presentations in order for leukocytes in flowing blood to interact rapidly and with high affinity to GMP-140 on activated platelets or endothelium.

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The three members of the selectin receptor family recognize a common carbohydrate epitope, the sialyl Lewis(x) oligosaccharide

The Journal of Cell Biology ◽

10.1083/jcb.117.4.895 ◽

1992 ◽

Vol 117 (4) ◽

pp. 895-902 ◽

Cited By ~ 459

Author(s):

C Foxall ◽

SR Watson ◽

D Dowbenko ◽

C Fennie ◽

LA Lasky ◽

...

Keyword(s):

Adhesion Molecule ◽

Leukocyte Adhesion ◽

Structural Features ◽

Sialyl Lewis X ◽

Lewis X ◽

Leukocyte Adhesion Molecule ◽

Lectin Domain ◽

Calcium Dependent ◽

Carbohydrate Epitope ◽

Sialyl Lewis

The selectins (lectin-EGF-complement binding-cell adhesion molecules [LEC-CAMs]) are a family of mammalian receptors implicated in the initial interactions between leukocytes and vascular endothelia, leading to lymphocyte homing, platelet binding, and neutrophil extravasation. The three known selectins, L-selectin (leukocyte adhesion molecule-1 [LECAM-1]), E-selectin (endothelial-leukocyte adhesion molecule-1 [ELAM-1]), and P-selectin (GMP-140) share structural features that include a calcium-dependent lectin domain. The sialyl Lewis(x) carbohydrate epitope has been reported as a ligand for both E- and P-selectins. Although L-selectin has been demonstrated to bind to carbohydrates, structural features of potential mammalian carbohydrate ligand(s) have not been well defined. Using an ELISA developed with a sialyl Lewis(x)-containing glycolipid and an E-selectin-IgG chimera, we have demonstrated the direct binding of the L-selectin-IgG chimera to sialyl Lewis(x). This recognition was calcium dependent, and could be blocked by Mel-14 antibody but not by other antibodies. Recognition was confirmed by the ability of cells expressing the native L-selectin to adhere to immobilized sialyl Lewis(x). These data suggest that the sialyl Lewis(x) oligosaccharide may form the basis of a recognition domain common to all three selectins.

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Glycan engineering using synthetic ManNAc analogues enhances sialyl-Lewis-X expression and adhesion of leukocytes

10.1101/2021.12.22.473788 ◽

2021 ◽

Author(s):

Anam Tasneem ◽

Shubham Parashar ◽

Tanya Jain ◽

Simran Aittan ◽

Jyoti Rautela ◽

...

Keyword(s):

Cell Adhesion ◽

Inflammatory Diseases ◽

Leukocyte Adhesion ◽

Fold Increase ◽

Sialyl Lewis X ◽

Lewis X ◽

Sialyl Lewis ◽

Acute Myeloid Leukemia Cells ◽

Coated Surfaces ◽

Cell Cell

Cell surface glycans, depending on their structures and dynamic modifications, act as the first point of contact and regulate cell-cell, cell-matrix, and cell-pathogen interactions. Particularly, the sialyl-Lewis-X (sLeX, CD15s) tetrasaccharide epitope, expressed on both glycoproteins and gangliosides, participates in leukocyte extravasation via interactions with selectins expressed on endothelial cells, lymphocytes, and platelets (CD62-E/L/P). Neutrophils carrying sLeX epitopes are thought to be responsible for chronic inflammatory diseases resulting in plaque formation and atherosclerosis. Intense efforts have been devoted to the development of sLeX mimetics for inhibition of cell adhesion. On the other hand, dysregulated expression of sLeX and poor extravasation are the major underlying causes of leukocyte adhesion deficiency-II (LAD-II) disorders that result in frequent infections and poor immune response. We hypothesized that metabolic processing of peracetyl N-(cycloalkyl)acyl-D-mannosamine derivatives, through the sialic acid pathway, might result in the expression of sialoglycans with altered hydrophobicity which in-turn could modulate their binding to endogenous lectins, including selectins. Herein, we show that treatment of HL-60 (human acute myeloid leukemia) cells with peracetyl N-cyclobutanoyl-D-mannosamine (Ac4ManNCb), at 50 microM for 48 h, resulted in a robust three to four fold increase in the binding of anti-sLeX (CSLEX1) antibody and enhanced cell adhesion to E-selectin coated surfaces; while the corresponding straight-chain analogue, peracetyl N-pentanoyl-D-mannosamine (Ac4ManNPent), and peracetyl N-cyclopropanoyl-D-mannosamine (Ac4ManNCp) both resulted in 2.0-2.5fold increase compared to controls. The ability to enhance sLeX expression using small molecules has the potential to provide novel opportunities to address challenges in the treatment of immune deficiency disorders.

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CD62 and endothelial cell-leukocyte adhesion molecule 1 (ELAM-1) recognize the same carbohydrate ligand, sialyl-Lewis x.

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.88.14.6224 ◽

1991 ◽

Vol 88 (14) ◽

pp. 6224-6228 ◽

Cited By ~ 309

Author(s):

M. J. Polley ◽

M. L. Phillips ◽

E. Wayner ◽

E. Nudelman ◽

A. K. Singhal ◽

...

Keyword(s):

Endothelial Cell ◽

Adhesion Molecule ◽

Leukocyte Adhesion ◽

Sialyl Lewis X ◽

Lewis X ◽

Leukocyte Adhesion Molecule ◽

Sialyl Lewis ◽

Carbohydrate Ligand

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Fluorinated per-acetylated GalNAc metabolically alters glycan structures on leukocyte PSGL-1 and reduces cell binding to selectins

Blood ◽

10.1182/blood-2009-07-231480 ◽

2010 ◽

Vol 115 (6) ◽

pp. 1303-1312 ◽

Cited By ~ 46

Author(s):

Dhananjay D. Marathe ◽

Alexander Buffone ◽

E. V. Chandrasekaran ◽

Jun Xue ◽

Robert D. Locke ◽

...

Keyword(s):

Inflammatory Diseases ◽

Leukocyte Adhesion ◽

Human Leukocyte ◽

Sialyl Lewis X ◽

Metabolic Inhibitor ◽

Growth Media ◽

Cell Binding ◽

Lewis X ◽

Sialyl Lewis

Abstract Novel strategies to control the binding of adhesion molecules belonging to the selectin family are required for the treatment of inflammatory diseases. We tested the possibility that synthetic monosaccharide analogs can compete with naturally occurring sugars to alter the O-glycan content on human leukocyte cell surface selectin-ligand, P-selectin glycoprotein ligand-1 (PSGL-1). Resulting reduction in the sialyl Lewis-X–bearing epitopes on this ligand may reduce cell adhesion. Consistent with this hypothesis, 50μM per-acetylated 4F-GalNAc added to the growth media of promyelocytic HL-60 cells reduced the expression of the cutaneous lymphocyte associated-antigen (HECA-452 epitope) by 82% within 2 cell doubling cycles. Cell binding to all 3 selectins (L-, E-, and P-selectin) was reduced in vitro. 4F-GalNAc was metabolically incorporated into PSGL-1, and this was accompanied by an approximately 20% reduction in PSGL-1 glycan content. A 70% to 85% reduction in HECA-452 binding epitope and N-acetyl lactosamine content in PSGL-1 was also noted on 4F-GalNAc addition. Intravenous 4F-GalNAc infusion reduced leukocyte migration to the peritoneum in a murine model of thioglycolate-induced peritonitis. Thus, the compound has pharmacologic activity. Overall, the data suggest that 4F-GalNAc may be applied as a metabolic inhibitor to reduce O-linked glycosylation, sialyl Lewis-X formation, and leukocyte adhesion via the selectins.

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Induction of superoxide anion production from monocytes and neutrophils by activated platelets through the P-selectin-sialyl Lewis X interaction

Journal of Leukocyte Biology ◽

10.1002/jlb.56.5.583 ◽

1994 ◽

Vol 56 (5) ◽

pp. 583-587 ◽

Cited By ~ 59

Author(s):

Tsutomu Tsuji ◽

Kisaburo Nagata ◽

Junzo Koike ◽

Naoko Todoroki ◽

Tatsuro Irimura

Keyword(s):

Superoxide Anion ◽

Sialyl Lewis X ◽

Lewis X ◽

Superoxide Anion Production ◽

Sialyl Lewis ◽

Activated Platelets ◽

Anion Production

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Discordant expression of selectin ligands and sialyl Lewis x–related epitopes on murine myeloid cells

Blood ◽

10.1182/blood-2002-06-1799 ◽

2002 ◽

Vol 100 (13) ◽

pp. 4485-4494 ◽

Cited By ~ 38

Author(s):

Marcin M. A. Kobzdej ◽

Anne Leppänen ◽

Vishwanath Ramachandran ◽

Richard D. Cummings ◽

Rodger P. McEver

Keyword(s):

Monoclonal Antibodies ◽

Sialic Acid ◽

Fluid Phase ◽

Sialyl Lewis X ◽

Lewis X ◽

Intact Cells ◽

Sialyl Lewis ◽

Selectin Ligands ◽

Low Levels ◽

Myeloid Leukocytes

Murine leukocytes are thought to express α2-3-sialylated and α1-3-fucosylated selectin ligands such as sialyl Lewis x (sLex), although monoclonal antibodies (mAbs) to sLex or Lex reportedly do not bind to murine leukocytes. We observed that P- and E-selectin bound to pronase-sensitive ligands on murine monocytic WEHI-3 cells and murine neutrophils, indicating that the ligands for both selectins are glycoproteins. CSLEX-1, HECA-452, and other widely used mAbs to sLex and Lex did not bind to WEHI-3 cells and bound at very low levels to murine neutrophils. Only the anti-sLex mAbs 2H5 and KM93, which also recognize nonfucosylated glycans, bound to WEHI-3 cells. 2H5 and KM93 bound to pronase-resistant structures, indicating that the mAbs did not identify selectin ligands. Treatment of WEHI-3 cells with glycosidases or chlorate demonstrated that sialic acid modifications, α1-3-galactosylation, or sulfation did not mask epitopes for mAbs to sLex or Lex. Compared to human promyelocytic HL-60 cells, WEHI-3 cells and murine neutrophils expressed low α1-3-fucosyltransferase activities. Consistent with very low endogenous fucosylation, forced fucosylation of intact WEHI-3 cells or murine neutrophils by exogenous α1-3-fucosyltransferase FTVI and GDP-fucose created many new epitopes for anti-sLexmAbs such as HECA-452 and CSLEX-1. Nevertheless, forced fucosylation of intact cells did not significantly augment their ability to bind to fluid-phase P- or E-selectin or to roll on immobilized P- or E-selectin under flow. These data suggest that murine myeloid leukocytes fucosylate only a few specific glycans, which interact preferentially with P- and E-selectin.

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